The immediate objective of the proposed project is to purify ectopic hCG-beta from culture fluids of the DoT and CaSKi cervical cancer cell lines, neither of which secretes significant amounts of intact hCG-alpha. The purification process will utilize affinity columns of anti-hCG linked to Sepharose, concanavalin A linked to Sepharose, gel filtration, and DEAE ion-exchange chromatography. The purified ectopic beta subunit of hCG will be characterized by gel electrophoresis, molecular sieve chromatography, activity in rat uterine weight bioassay and rat testicular receptor assay before and after combination with ectopic or eutopic hCG-alpha, and immunologic behavior in homologous radioimmunoassays for hCG and hCG-beta. It is hoped that this project will establish a processing system for the continuing purification of ectopic hCG-beta for future collaborative investigation, and a comparison of ectopic hCG-beta with eutopic hCG-beta. The long-range goal of this research is to develop a process for purifying small quantities of metabolic product (ectopic hCG-beta) from large volumes of serum-containing tissue culture fluids that are otherwise discarded routinely.